Sequence identification and in silico characterization of novel thermophilic lipases from Geobacillus species

In this study, we cloned and sequenced Lip21 and Lip33 genes fromGeobacillus sp. GS21 andGeobacillus sp. GS33, then we in silico and experimentally analyzed the encoded lipases. For this purpose, Lip21 and Lip33 were cloned, sequenced, and their amino acid sequences were investigated for determination of biophysicochemical characteristics, evolutionary relationships, and sequence similarities. 3D models were built and computationally affirmed by various bioinformatics tools, and enzyme-ligand interactions were investigated by docking analysis using six ligands. Biophysicochemical property of Lip21 and Lip33 was also determined experimentally and the results demonstrated that they had similar isoelectric point (pI) (6.21) andTm (75.5 °C) values asTm was revealed by denatured protein analysis of the circular dichroism spectrum and pI was obtained by isoelectric focusing. Phylogeny analysis indicated that Lip21 and Lip33 were the closest to lipases fromGeobacillus sp. SBS-4S andGeobacillus thermoleovorans, respectively. Alignment analysis demonstrated that S144 –D348–H389 was catalytic triad residues in Lip21 and Lip33, and enzymes possessed a conserved Gly-X-Ser-X-Gly motif containing catalytic serine. 3D structure analysis indicated that Lip21 and Lip33 highly resembled each other and they were α/β hydrolase-fold enzymes with large lid domains. BAN ΔIT analysis results showed that Lip21 and Lip33 had higher thermal stability, compared to other thermostableGeobacillus l...
Source: Biotechnology and Applied Biochemistry - Category: Biochemistry Authors: Tags: ORIGINAL ARTICLE Source Type: research